Na Hee Sam, Park Mi Hee, Song Yu Ri, Kim Seyeon, Kim Hyung-Joon, Lee Ju Youn, Choi Jeom-Il, Chung Jin
Department of Oral Microbiology, School of Dentistry, Pusan National University, Yangsan, Korea.
Department of Oral Physiology, School of Dentistry, Pusan National University.
J Periodontol. 2016 Sep;87(9):e173-82. doi: 10.1902/jop.2016.160033. Epub 2016 May 31.
Periodontitis is a chronic inflammatory disease resulting from an inflammatory response to subgingival plaque bacteria, including Porphyromonas gingivalis. MicroRNA (miRNA) is a current focus in regulating the inflammatory processes. In this study, the inflammatory miRNA expression in gingival tissues of patients with periodontitis and of healthy individuals is compared, and its role in regulating the inflammatory response is examined.
Gingival tissues from patients with periodontitis and healthy individuals were collected for miRNA microarray. THP-1 and CA9-22 cells were challenged with P. gingivalis, and miRNA expression was determined by real-time polymerase chain reaction. Target genes for miRNA were predicted using TargetScanHuman database, and miRNA gene expressions were reviewed using public databases. For the functional study, THP-1 cells were transfected with a miRNA-128 mimic, and target gene expression was compared with THP-1 cells challenged with P. gingivalis. For the tolerance test, THP-1 cells transfected with miRNA-128 mimic were treated with phorbol 12-myristate 13-acetate (PMA) or paraformaldehyde (PFA)-fixed Escherichia coli. Tumor necrosis factor (TNF)-α production was determined by enzyme-linked immunosorbent assay, and mitogen-activated protein kinase (MAPK) protein phosphorylation was determined by Western blot.
Gingival tissues from patients with periodontitis showed increased expression of miRNA-128, miRNA-34a, and miRNA-381 and decreased expression of miRNA-15b, miRNA-211, miRNA-372, and miRNA-656. THP-1 cells and CA9-22 cells challenged with P. gingivalis showed increased miRNA-128 expression. Among the predicted miRNA-128 target genes, several genes that are involved in MAPK signaling pathway showed similar gene expression pattern between P. gingivalis challenge and miRNA-128 mimic transfection. In THP-1 cells transfected with miRNA-128 mimic, TNF-α production was lower, and phosphorylation of p38 was inhibited when challenged with PMA or PFA-fixed E. coli.
miRNA-128 may be involved in mitigating the inflammatory response induced by P. gingivalis in periodontitis.
牙周炎是一种由对龈下菌斑细菌（包括牙龈卟啉单胞菌）的炎症反应引起的慢性炎症性疾病。微小RNA（miRNA）是目前调节炎症过程的研究热点。在本研究中，比较了牙周炎患者和健康个体牙龈组织中炎症相关miRNA的表达，并研究了其在调节炎症反应中的作用。
收集牙周炎患者和健康个体的牙龈组织进行miRNA芯片检测。用牙龈卟啉单胞菌刺激THP-1细胞和CA9-22细胞，通过实时聚合酶链反应测定miRNA表达。使用TargetScanHuman数据库预测miRNA的靶基因，并通过公共数据库检索miRNA基因表达情况。为进行功能研究，用miRNA-128模拟物转染THP-1细胞，并将靶基因表达与用牙龈卟啉单胞菌刺激的THP-1细胞进行比较。为进行耐受性试验, 用佛波酯12-肉豆蔻酸酯13-乙酸酯（PMA）或多聚甲醛（PFA）固定的大肠杆菌处理转染了miRNA-128模拟物的THP-1细胞。通过酶联免疫吸附测定法测定肿瘤坏死因子（TNF）-α的产生，并通过蛋白质印迹法测定丝裂原活化蛋白激酶（MAPK）的磷酸化。
牙周炎患者的牙龈组织中miRNA-128、miRNA-34a和miRNA-381表达增加，而miRNA-15b、miRNA-211、miRNA-372和miRNA-656表达降低。用牙龈卟啉单胞菌刺激的THP-1细胞和CA9-22细胞中miRNA-128表达增加。在预测的miRNA-128靶基因中，几个参与MAPK信号通路的基因在牙龈卟啉单胞菌刺激和miRNA-128模拟物转染之间表现出相似的基因表达模式。在用miRNA-128模拟物转染的THP-1细胞中，当用PMA或PFA固定的大肠杆菌刺激时，TNF-α的产生较低，且p38的磷酸化受到抑制。
miRNA-128可能参与减轻牙周炎中牙龈卟啉单胞菌诱导的炎症反应。
